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承辦人： 謝秀梅 聯絡電話： 77346354 日期：102 年 3 月 4 日

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計 畫 名 稱 出席 2012 退化性神經疾病研討會發表論文

出 國 人 員

姓名 服務單位 職稱

謝秀梅 生命科學系 教授

出 國 地 點 墨西哥 坎昆

出 國 期 間 自 101 年 11 月 27 日起至 101 年 12 月 1 日止

承辦單位

核章

人事室

會核意見

校長核示

報 告 內 容

一、目的：2012 退化性神經疾病研討會(2012 Neurodegeneration Conference)

二、過程：

The 2012 Neurodegeneration Conference was taken placed on Nov. 27–Dec. 1, 2012, Cancun, Mexico. It was organized by Zing Conference, focused on the fields of neurodegenerative diseases, diagnosis and therapy. This meeting provided a chance for scientists to discuss and learn about all the important international breakthrough developments in whole genome genetic screening, drug discovery and other new therapeutics. The meeting thus can stimulate new and productive interactions among basic and clinical research groups involved in this field. The 5-day meeting was hold in the Occidental Grand Xcaret, Cancun, Mexico. There were total 14 Sessions, and 3-5 talks in each Session. The poster section was hold at the second day afternoon. It has more than 100 participants came from15 different countries to enjoy the updated knowledge and techniques in the study of updated knowledge in neurodegeneration. My poster presentation was entitled「Identification of HDAC inhibitors as potential therapy for polyQ-mediated spinocerebellar ataxia type 17」.

三、心得：

There were many impressive presentations during this conference. I have organized some of the outlines from the presentations and present it in this report. The summary reveals the progress of the science and medicine in neurodegenerative diseases in these days, which I am thinking could be our next approach in disease mechanism studies. In the Session 1: Alzheimer's disease, an overview. Eric Karran reviewed the “Developing drugs for Alzheimer’s disease (AD): past, present and future”. Nibaldo Inestrosa talked about “Regulation of the Neuronal Function by Wnts”. These two talks correlated to the research projects we are focusing. We have learned that there is no effect treatment for AD, and antibody approach is the most popular strategy toward the treatment. Wnt signaling pathway is highly associated with the neurodegeneration and could be a potential therapeutic target of neurodegenerative diseases.

Session 2: Genetics of Neurodegenerative diseases. Dr. John Hardy: Genetic analysis of neurodegeneration. He mentioned the 3 points about the whole genome genetics: (1) Powerful technology to find genetic risk; (2) risk is predictable; (3) late onset can be predisposed by heterozygous variants. He also talked about their recent 2

projects, recessive FTD and recessive AD Turkish population, and pathology confirmed Caucasian AD. Dr. Olga Corti: Novel molecular mechanisms uderlying the mitochondrial quality control functions of the PINK1/Parkin pathway. The Parkin is recruited to mito after mito loss the import efficiency and Parkin is critical for mitophagy-Ub-proteosome pathway. Val Moskvina from Cardiff University, UK, talked about the GWAS among AD/control. IGAP 4 consotrtia collaboration with 18600 cases and 41370 contl. They identified 12 new genes associated with AD, 8 of the 12 genes were from GWAS, and the other 4 genes were from gene-based analysis. Rovelet-Lecrux: Involvement of novel genes in autosomal dominant early-onset AD and reassessment of the role of APOE. With exon sequencing, they studied SOLR1 gene in 7/29 ADEOAD and found 2 unknown variation and made the knockout (KO) mice in which Aβ is increased. Minerva Carrasquillo from Mayo clinic talked the Brain Expression Genome-Wide Association Study (eGWAS) Identifies Human Disease-Associated Variants Combined disease and express GWAS. 9 novel loci were identified, including MS4a4A, CLU and ABCA7.

Martine Barkats: AAV gene transfer to the CNS: application to gene therapy of Spinal Muscular Atrophy. They presented a promising therapeutic results, which even surpassed those reported after IV or IM injection, were also obtained following intracerebroventricular (ICV) and co-ICV/IV injections of similar vectors in SMNΔ7 mice. Jean-Daniel Abraham: Specific detection of N-truncated Abeta peptides in complex fluids. They found higher Aβ11-40 in AD not Aβ17-40, so they can improve AD diagnostis by specifically measuring Aβ11-40 peptide, highlighting the possible role of β-secretase dysregulation in AD physiopathology.Mark Cookson talked about the PD gene-Leucine-rich repeat kinase 2 (LARRK2). They functional assayed the LARRK2 mutants which impaired the function in endocytosis, and identified the LARRK2 interacting protein GAK Ugrumov’s talk topic is “Parkinson’s disease and hyperprolactinemia: new insight into pathogenesis, diagnostics and therapy”. He talked about the pathomechanism and the related diagnosis of PD. Selina Wray has made the iPS models of frontotemporal dementia caused by mutation in Tau and a non-coding hexanucleotide repeat expansion in C9ORF72 , which is also a gene related to ALS.

Session 5: Molecular cross-talk between prion and Abeta Frédéric Checler: Proteolytic events govern a molecular cross-talk between prions and Alzheimer's disease pathologies. α-secretase cut both APP and PrP (regulated TACE, constitutive ADAM10). The N1 and sAPPu protect cells against p53-dep apoptosis, and the protection is abolished in LRP1-deficient cells. Nigel Hooper: Prion protein as a receptor for amyloid-β oligomers – implications for Alzheimer’s disease.

Prion regulates the production and action of Aβ. Oligomer Aβ, the ADDLs (5-6 nm) was used in their study. Two antibodies (Ab), A11 Ab against pre-fibrill oligomers and OC Ab against fibril oligomers were used to detect the Aβ loads. They found Aβ oligomer require PrPc to mediate cytotoxicity and activation of Fyn kinase.

LRP1 and PrP associated for Aβ oligomer to bind to trigger the toxicity. Aβ oligomer endocytose with PrP to lysosome. EGCG and resverval reduced the binding of Aβ oligomer bind to PrP cells and the toxicity. In addition, PrP facilitate Zn uptake into the neuron via AMPA receptor promote Aβ formation. This was confirmed by PrP knockdown (KD) which reduce Zn uptake. Vilma Martins: Prion protein (PrP)-dependent neurotrophic signals are potentiated by astrocyte-released endosomal microvesicles carrying the co-chaperone STI1. STI1-PrP binding induce neuronal survival and differentiation (via ERK pathway) /Modulate protein synthesis (via mTOR). PrPc is a scaffold and STI1 is detected in the conditioned media of astrocyte not neuron (STI1 is secreted by astrocytes). The STI1 media induce neuron usurvival. STI1 is secreted in vesicles (exosomes) that are heterogeneous in size and as a neurotrophic factor using PrP as receptor. Aβ oligomers (AβO) reduced the synatophysin expression (exp) and rescued by STI1 and in PrP deficient cells AβO is non-toxic, therefor, using a ligand to block the interaction of AβO and PrP is potential in AD treatment. Marco Prado: The Prion Protein Partner, STI1, is involved in embryonic development, cellular stress and protects neurons against Abeta-induced cell death. PrP is GPI-anchored extracellular scaffolding (square dancer lady change parterners during the dance was used as an example in the talk). Ac increase cell surface PrP and KO mice is e10.5 lethal. Oxygen-glucose depletion reduced the STI exp. Aβ bind to PrP was identified by using SPR and the interaction was competed by STI1.

Herbert Tabor Prize Lecture was given to James Duce in University of Leeds (UK). Tau KO cells have impaired ion influx and the ion accumulation in tau KO neurons. APP ferroxidase is reduced in AD cortical tissue and aged Tg2578 mice. APP ferroxidase activity partially impaired by Aβ bound Zn in AD. In addition, loss of tau is associated with PD. Overexp of APP protect against MPTP toxicity

Session 6: AD and the cholinergic system. Hermona Soreq: MicroRNAs in the interface between inflammation and neurodegeneration. miRNA132 interact with AchE 3’UTR and miR608 target AChE and CDC42. Animals with more CDC42 show anxiety. hnRNP A/B KD impair learning and memory. Furthermore, miR211 increased in AD while miR132 reduced in AD. Vania Prado:Consequences of cholinergic dysfunction in the cortex and hippocampus: highlights from VAChT-deficient mice. VAChT-deficient mice die at E18.5 and increase

motoneurons.Forebrain KO with Six3-cre mice is hyperactive, reversal learning impairment, attention deficiet (increased omission) vigilance.

Session 7: APP metabolism & regulation I. Sangram Sisodia: Presenilin function in health and disease. PS1 facilitate r-secretase processing of APP. Nicastrin a gatekeeper for r-secretase. Conditional KO of PS1dE9 in HNPCs in hippo and forebrain rescue AHNPC proliferation, not the maturation. Edward Parkin: The amyloid precursor protein and proteolysis; molecular overlaps in Alzheimer’s lower prevalent to develop of cancer in AD patients was identified. P53 repress APP promoter activity and APP exp in several cancers were found. Ir20a inhibit proliferation and invention of ovarien cancer. APP protects DU145 cells against Coppernediate toxicity.

Session 8: APP metabolism & regulation II. Guojun Bu: ApoE and apoE receptors in brain lipid metabolism, synaptic functions and clearance of amyloid-beta. He talked about why ApoE4 is a strong risk factor of AD. Soluble ApoE are decreased and insoluble are increased in AD. cKO of he receptor LRPI (CamK2-cre) in an age-dep study –impairment in lipid metabolism. NMDAR1 and GluR1 reduced GFAP, increased clasping, rotard latency decreased, neuronal cell loss, synapse lose, impair insulin signaling and glu metabolism. Aβ is endocytosis in ER (ApoE) to lysosome and part is recycled. E3 is better than E4 in the transferring. Tobias Hartmann: AICD and Aβ-regulator and target of neuronal lipid metabolism. DHA reduce secretase activity and further decrease Aβ production.

Session 9: Biomarkers & Therapeutics. Dale Schenk: Tau based models and approaches for the treatment of Neurodegenerative diseases. Tau kinase inhibition and tau progression inhibition are potential treatment strategy for AD. JPNL3 TauP301S mice have reproducible phenotypes and used for animal model in their studies. MAPK inhibitor SRN-003-556 10-20mg/kg, PR (GSK3 inhibitor) 1mg/kg, Epothilone 0.3-1 mg/kg were applied to the mice to determine their therapeutic effects. Nicolas Sergeant: In vivo therapeutic strategies against tau pathology: From exercise to immunotherapy. Tau aggregates occur in aging and more than 20 disorders. Voluntary exercise wheels for 40 weeks on mice improved the phenotypes of transgenic mice. David Allsop: Development of peptide-inhibitor nanoparticles (PINPs) as inhibitors of AβO formation. Amyloid cascade hypothesis is a trigger oxidative stress and tau phosphorylation and microglia activation further cause the cell death. Peptide NH2-RG-KLVFF-GR-NH2 (OR2) was not stable, and the RI-OR2 (reverse inverted) was more stable and blocked the fibril formation identified by Thio-T assay. RI-OR2-TAT affinity was shown increased, APP/PS1 mice 1 hr after inj Aβ found peptide bind to Aβ and microglia (Iba1 staining). The in

vivo assay is not conducted yet.

Session 10: Beyond classical therapeutic approaches. Tom Boyer: Mediator as a transducer of Amyloid Precursor Protein-dependent nuclear signaling. AICD fragment of APP goes to nuclei to regulate target genes, NEP and P53. MED12 is a hub for many signaling pathway and found several genetic variants are associated with many diseases. Target genes are increased in AD and PD patients. Marcela Karpuj (Bio AZ2 founder and CEO Isrel): A novel method for membrane protein regulation in cell culture. PS-DNA degraded PrP transiently. The N-term is required for degradation and degradation only occurred on plasma membrane. Gemma Casadesus Smith: Targeting Metabolic Hormone Dysregulation as a Treatment for AD. Leptinis is 270× more potent than insulin in reduction of tau phorsphorylation. Amylin was found reduced in AD and MCI patients. SAMP8 treatment after amylin improved cognition spatial memory, reduction of oxidative stress (HNE and Cox2 exp staining), HO-1 exp expression (WB), synapsin 1 in SAMP8 and APP PS1 mice. Combine treatment of leptin and amylin on SAMP8 mice showed reduced the latency in target quart time and BrdU staining in hippocampus. René Vidal: Targeting the UPR transcription factor XBP1 protects against Huntington’s disease in vivo. XBP1 KO HD (Q128) mice showed improvement in motor, neuron survival, DARPP32 through autophagy via inhibition of FoxO1.

Session 11: Amyloid lowering mechanisms I. Takaomi Saido: A new mouse model of Alzheimer's disease: Distinguishing facts from artifacts. We need a model overexp Aβ but not Ap. They created many different mice. 145F mice with Aβ42/40>30X. APP NL-F KI mice showed neurodegeneration in DG of hippocampus at 15mo by Th-S staining. NL-F x calpanstain KO mice showed no effect on life-span and no early lethality. Raphaelle Pardossi-Piquard: AICD: a key player in AD pathology? AICD is preferently produced by C99 and less from C83. Alpha-secretase is not essential for AICD production and AICD is pathogenic in vivo. C99 and AICD are early cause of AD. Natalia Nalivaeva: Amyloid-degrading enzymes as therapeutic targets in Alzheimer's disease. HDACi and VPA I NEP expression and activity reduced Aβ load (Th-S). HDAC3 is important in neuron and HDAC1 and 3 on the promoter of NEP.

Session 12: Amyloid lowering mechanisms II. Malcolm Leissring: Proteolytic degradation of the amyloid beta-protein: an historical perspective. Aβ degrading proteases CatD is a lysosomal protease. It works more efficiently in Aβ42 than Aβ40. The CatD KO mice showed insoluble Aβ was increased. Laura Morelli: Human Insulin-degrading enzyme (IDE) is a target of mitochondrial biogenesis pathway and modulates mitochondrial Aβ accumulation. IDE is a Zn-dep

metaloendoprotease with many targets: Aβ, insulin and AICD. Met42 is a cytosol form while Met1 is a mito form. Scott Miner: Enhancing amyloid β catabolism by intracerebral convection-enhanced delivery of neprilysin. NEP was shown increased in AD and aging and CEC2 was increased in AD. These are protective compensatory response.

Session 14: New tracks for innovative therapeutic approaches. Marco Racchi: Role of p53 in Alzheimer’s disease pathogenesis. They identified a defective activity of p53 in AD fibroblasts due to conformational changes in p53 tertiary structure. Unfolded p53 found in AD fibroblasts by exposure to Aβ 1-40 and Aβ 1-42. These unfolded p53 conformation may participate in AD development and considered as a marker of the early stage of AD. Andrea Leblanc: Caspase-6 activation in the hippocampus induces age-dependent memory impairment in mice and humans Active Caspase-6 is present in the NFT of AD brains and is as abundant in mild AD than moderate and severe AD. They found levels of ERC and CA1 Casp6 activity correlated with lower global cognitive scores, and episodic and semantic memory performance. Regression analyses revealed that Casp6 activity is the major contributor to lower episodic memory performance. A transgenic mouse that expresses Casp6 in the CA1 and memory deficits were observed with the Morris water maze and an object recognition task, occurred at 16-18 months of age.

From these seminars, I learned lots new technologies and knowledge. Some of them can be applied to our researches right away and we will try to work on it. I think the impact from this meeting can really promote our research works.

四、建議：

This meeting really provides an excellent opportunity for scientist to overview the advances and study development in neurodegenerative diseases. Participants focusing on different research field can also communicate about the experience in drug application to different diseases. Scientists and graduate students who are working on translational medicine are encouraged going to this meeting.

五、其它（自行決定是否附論文或相片…等資料）

Poster presentation